Method and apparatus for determining the rate of enzyme reaction

• US 4,224,405 A
• Filed: 04/26/1978
• Issued: 09/23/1980
• Est. Priority Date: 05/10/1977
• Status: Expired due to Term

• Current Assignee: Olympus Optical Corporation Limited (Olympus Corporation)
• Original Assignee: Olympus Optical Corporation Limited (Olympus Corporation)
• Inventors: Hijikata, Kazuo
• Primary Examiner(s): Warden, Robert J.
• Application Number:
  US05/900,149
• Time in Patent Office: 881 Days
• Field of Search: 195/103.5 R, 195/127, 195/139, 422/68, 356/409, 356/414, 356/436, 364/497, 364/498, 364/499, 364/500, 23/230 B, 435/3, 435/4, 435/288, 435/289, 435/290, 435/291, 435/808
• US Class Current: 435/3
• CPC Class Codes: G01N 21/272 for following a reaction, e.g. for determining photometrically a reaction rate (photometric cinetic analysis)

Abstract

The rate of enzyme reaction is determined by determining of the quantity of enzyme, and deriving the rate of reaction per unit time of a reaction solution having an absorbance which is proportional to the time. A temperature correction coefficient which is used during the determination of the enzyme quantity is derived by temperature information which is supplied by a temperature sensor disposed within the reaction solution to be determined. An apparatus used for the determination of the rate of reaction comprises a means for containing the reaction solution, a temperature sensor directly disposed within the solution, a photoelectric transducer element responsive to light transmitting through the solution to determine the optical density of the solution, a logarithmic conversion circuit which converts the signal from the element into a corresponding absorbance signal, and a coefficient conversion circuit which is responsive to the combination of the absorbance signal and temperature information from the temperature sensor to correct a temperature coefficient.

Claims

1. A method of determining the rate of an enzyme reaction in which an enzyme quantity is determined from the rate of a reaction in a reaction solution having an absorbance changing approximately linearly with the time;

said method comprising the steps of;

disposing a temperature sensor in the reaction solution for measuring the temperature thereof;

modifying the value of a temperature correction coefficient in accordance with temperature information from said sensor to correct for temperature changes in said reaction solution; and

using said value of said temperature correction coefficient to determine the rate of said enzyme reaction.

2. An apparatus for determining the rate of an enzyme reaction, comprising:

reaction solution container means for containing a quantity of a reaction solution and including a transparent portion which permits light to pass through said reaction solution contained therein;

a temperature sensor disposed in a position inside said container means and in direct contact with said reaction solution when said transparent cell contains said reaction solution for measuring the temperature thereof;

a photoelectric transducer element responsive to light which is passed through said solution;

a logarithmic conversion circuit for converting an electrical signal generated by said transducer element into a signal indicative of the absorbance of said reaction solution; and

a coefficient conversion circuit for modifying a temperature correction coefficient used in the determination of the enzyme quantity responsive to both said signal from said logarithmic conversion circuit and temperature information from said temperature sensor.

3. An apparatus according to claim 2 in which said temperature sensor comprises a thermistor having its surface provided with a coating of a water-repellent-material to prevent contamination.

4. An apparatus according to claim 2 in which said coefficient conversion circuit comprises:

an amplifier for amplifying said signal generated by said temperature sensor;

an analog-digital converter having a plurality of outputs for converting the output generated by said amplifier into corresponding digital signals;

a plurality of field effect transistors equal in number to said plurality of outputs and having their respective gate electrodes connected with respective individual ones of said outputs of aid analog-digital converter so as to be rendered conductive responsive to said digital signals, and

a second amplifier connected in series with each of said transistors and also receiving a signal generated by said logarithmic conversion circuit.

5. An apparatus according to claim 2, wherein said reaction solution containing means further comprises a transparent cell for containing said reaction solution.

6. An apparatus according to claim 5, wherein said reaction solution containing means further comprises a thermostatic block accommodating said transparent cell.

7. An apparatus according to claim 2, further comprising output printer means for printing data representative of the output of said coefficient conversion circuit.